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Volume 7, Issue 1 (Suppl) J Biotechnol Biomater
ISSN: 2155-952X JBTBM, an open access journal
March 20-21, 2017 Rome, Italy
&
15th World Congress on
2nd International Conference on
Biotechnology And Biotech Industries Meet
Enzymology and Molecular Biology
Enzymology & Mol. Biology 2017
Biotechnology Congress 2017
March 20-21, 2017
Continuous methods of fungal laccase concentration
Michal Blatkiewicz1, Anna Antecka1, Stanislaw Ledakowicz1 and Andrzej Gorak1, 2
1Lodz University of Technology, Poland
2Dortmund University of Technology, Germany
Statement of the Problem: Downstream processing of biological molecules is a very time- and energy-consuming task. One of the
major trends in contemporary biotechnology revolves around cost-effective and environment-friendly methods of concentration and
purification of bioproducts. Various novel downstream processing tactics are currently being investigated as alternatives to established
methods such as ultrafiltration and chromatography. The purpose of this research was to examine the feasibility of polyethylene
glycol-phosphate aqueous two phase systems (ATPS) and cetrimonium bromide-induced foam fractionation (FF) as methods for
Cerrena unicolor and Pleurotus sapidus laccase separation from culture supernatants. Both processes were investigated in batch and
continuous forms.
Methodology: The biosynthesis was performed in a 14-L bioreactor equipped with a set of sensors for process control. The filtered
supernatants were concentrated with the use of aqueous two-phase systems or foam fractionation. Batch ATPS experiments were
conducted in specially designed extraction flasks, and for continuous ATPS experiments, a mixer-settler unit (MSU) was used. FF
experiments were conducted in a special glass column equipped with air disperser and peristaltic pumps for liquid intake and outtake.
Laccase activity was determined by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) assay.
Findings: C. unicolor laccase showed greater affinity towards salt-rich phase with over 90% yields and partitioning coefficients up
to 2200. P. sapidus laccase showed strong affinity towards polymer rich-phase also with over 90% yields and full partitioning. MSU
experiments showed consistency with batch experiments within non-extreme phase ratio range. Foam fractionation effectiveness
depended strongly on pH and surfactant concentration, leading over 100 partitioning coefficient towards the foamate. Low gas and
liquid flow rates led to more effective concentration.
Conclusions: Aqueous two-phase extraction and foam fractionation are both effective alternatives to established downstream
processing methods for laccase concentration.
Biography
Michal Blatkiewicz has done his Master of Engineering Technology in the field of Chemical and Process Engineering from Cracow University. Currently, he is a PhD
student at Lodz University of Technology, Faculty of Process and Environmental Engineering, where he is also employed as a Scientific Project Contractor. During
his PhD studies, he has done four internships at Dortmund University of Technology. His scientific scope includes primarily fungal cultures and enzymes, and also
downstream processing of biological molecules. Currently, he is a part of a research team working on a project concerning continuous processes of biosynthesis,
concentration, and purification of fungal laccases, in which he focuses mostly on novel downstream processing methods, such as aqueous two-phase extraction
and foam fractionation.
Michal Blatkiewicz et al., J Biotechnol Biomater 2017, 7:1(Suppl)